| Objective:To investigate the expression level of long non-coding RNA(lncRNA)RP11-363E7.4 in gastric cancer(GC)tissues,adjacent normal tissues and GC cell lines,the correlation between the expression level of lncRNA RP11-363E7.4 and the clinicopathological characteristics of GC patients,the effects of lncRNA RP11-363E7.4 on proliferation,apoptosis,migration and invasion of GC cells,and the underlying potential molecular mechanism.Methods:1.The expression level of lncRNA RP11-363E7.4 in 82 paired of GC tissues and adjacent normal tissues were detected by qRT-PCR.Then the correlation between lncRNA RP11-363E7.4 and the clinicopathological characteristics of GC patients were analyzed.2.The expression level of lncRNA RP11-363E7.4 in 3 human GC cell lines(SGC-7901,BGC-823,MGC-803)and human normal gastric mucosa epithelial cell(GES-1)were detected by qRT-PCR.Followed by the construction of a stable overexpression cell line of lncRNA RP11-363E7.4.Then the transfection efficiency of lncRNA RP11-363E7.4 was detected by qRT-PCR.3.The effects of lncRNA RP11-363E7.4 on the proliferation,apoptosis,migration and invasion of gastric cancer cells were detected by CCK-8 cell proliferation assay,colony formation assay,flow cytometry,scratch wound healing assay and Transwell migration and invasion assay.4.The expression level of proteins related to apoptosis and EMT in lncRNA RP11-363E7.4 overexpressed GC cells were detected by western blot assay to explore the underlying potential molecular mechanisms.Results:1.The expression level of lncRNA RP11-363E7.4 in GC tissues was significantly lower than that in the corresponding normal tissues(p <0.05).The expression level of lncRNA RP11-363E7.4 was correlated with T stage(p<0.05),but not with age,gender,Borrmann classification,tumor size,M stage and N stage(all p>0.05).2.The expression level of lncRNA RP11-363E7.4 in three human GC cell lines was significantly lower than that in GES-1 cell line,and the expression level in SGC-7901 and MGC-803 cell lines were lower than that in BGC-823 cell line.LncRNA RP11-363E7.4 was up-regulated for 32.49 times in SGC-7901 cells(p<0.05)and for 33.92 times in MGC-803 cells(p<0.05).3.Overexpression of lncRNA RP11-363E7.4 can inhibit proliferation and induce apoptosis of GC cells.Moreover,the protein expression level of Caspase-3,Bax and p53 were increased and that of Bcl-2 was decreased in the lncRNA RP11-363E7.4 overexpressed group compared to the negative control group.4.Overexpression of lncRNA RP11-363E7.4 can inhibit migration and invasion of GC cells.Moreover,the protein expression level of Ecadherin was increased and that of N-cadherin,Vimentin,Slug and ?-catenin was decreased in the lncRNA RP11-363E7.4 overexpressed group compared to the negative control group.Conclusion:1.The expression level of lncRNA RP11-363E7.4 was significantly downregulated in gastric cancer tissues and cell lines than that of the corresponding normal ones,and the low expression level of RP11-363E7.4 was associated with T stage of tumor.2.LncRNA RP11-363E7.4 can inhibit proliferation,migration and invasion and induce apoptosis of gastric cancer cells to play a role of tumor suppression.3.LncRNA RP11-363E7.4 can influence the expression level of apoptosis related proteins and EMT related protein in gastric cancer cells,which may be related to its tumor suppression effect in gastric cancer cells. |
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